Methods or protocols for the detection and quantitation of specific nucleic acid sequences including nucleic acid purification, denaturation and the like are well known in the field of research molecular biology. Much of the work in the field of thermal renaturation of DNA has been performed by Marmur and Doty, (1961) J. Mol. Biol. 3, 585 wherein comparisons are made between DNA and renatured DNA showing the reproducibility of the helix-coil transition and the reformation of the same secondary structure.
A technique for the immobilization of denatured DNA to a solid substrate, such as a nitrocellulose filter, and subsequent renaturation or hybridization by a complimentary RNA has been reported by Gillespie and Siegelman (19-5), J. Mol. Biol. 12, 829. Such technique was further modified and extended by Denhardt, (1966) Biochemical and Biophysical Research Communications 23, No. 5, 641, to detect complimentary denatured DNA sequences with the use of a labeled DNA probe. A method for the detection of Epstein-Barr viral DNA in lymphoid cell lines is disclosed by Braudsma and Miller, (1980) Proc. Natl. Acad. Sci. U.S.A.
One clinical method available to determine infectivity of a suspect human serum for a Hepatitus-B virus, presently requires in vivo inoculation of the suspect human serum into a chimpanzee, although many procedures are available to detect products of such a suspect viral deoxyribonucleic acid. The use of chimpanzees is costly and time consuming, whereas the detection of such products is indirect. The detection of other pathogenic agents present like problems of cost, unreliability, need for invasion sampling and the like.